摘要
目的探讨长链非编码RNA 01106(LINC01106)调节微小RNA-372-3p(miR-372-3p)/组蛋白甲基转移酶2(SMYD2)轴对肝癌细胞增殖、迁移和侵袭的影响。方法实时荧光定量PCR(qRT-PCR)检测肝癌组织和细胞(Huh-7、HepG2、SMMC-7721)中LINC01106、miR-372-3p、SMYD2的表达水平;将肝癌SMMC-7721细胞分为control组、si-LINC01106组、si-NC组、si-LINC01106+miR-372-3p inhibitor组、si-LINC01106+inhibitor-NC组;检测5组SMMC-7721细胞中LINC01106、miR-372-3p、SMYD2的表达水平;MTT法与Edu染色检测SMMC-7721细胞增殖;划痕实验检测SMMC-7721细胞迁移;Transwell小室检测SMMC-7721细胞侵袭;WB方法检测PCNA、MMP-2、MMP-9、SMYD2蛋白表达;双萤光素酶报告基因实验证实LINC01106或SMYD2与miR-372-3p的关系。结果LINC01106和SMYD2在肝癌组织和细胞系中显著上调,miR-372-3p在肝癌组织和细胞系中显著下调(P<0.05)。与control组、si-NC组比较,si-LINC01106组SMMC-7721细胞LINC01106 mRNA、SMYD2 mRNA、OD490、Edu阳性细胞率、细胞划痕愈合率、细胞侵袭数、PCNA蛋白、MMP-9蛋白、MMP-2蛋白、SMYD2蛋白表达下降,miR-372-3p表达增高(P<0.05);与si-LINC01106+inhibitor-NC组、si-LINC01106组比较,si-LINC01106+miR-372-3p inhibitor组SMMC-7721细胞SMYD2 mRNA、OD490、Edu阳性细胞率、细胞划痕愈合率、细胞侵袭数、PCNA蛋白、MMP-9蛋白、MMP-2蛋白、SMYD2蛋白表达增高,miR-372-3p表达下降(P<0.05)。LINC01106靶向负调控miR-372-3p表达,SMYD2被miR-372-3p靶向下调。结论LINC01106敲低可能通过靶向miR-372-3p/SMYD2轴抑制SMMC-7721细胞迁移、侵袭和增殖。
Objective To investigate the impacts of long non-coding RNA 01106(LINC01106)on the proliferation,migration,and invasion of liver cancer cells by regulating the microRNA-372-3p(miR-372-3p)/SET and MYND domain containing protein 2(SMYD2)axis.Methods Real-time fluorescence quantitative PCR(qRT-PCR)was applied to detect the expression levels of LINC01106,miR-372-3p,and SMYD2 in liver cancer tissues and cell lines(Huh-7,HepG2,SMMC-7721).SMMC-7721 cells were induced with blank control,or transfected with si-LINC01106,si-NC,si-LINC01106+miR-372-3p inhibitor or si-LINC01106+inhibitor-NC,followed by measurements of LINC01106,miR-372-3p and SMYD2 levels.Cell proliferation was detected by MTT assay and EdU staining.Cell migration and invasion were examined by wound healing assay and Transwell assay,respectively.Western blot(WB)was applied to detect the protein expressions of PCNA,MMP-2,MMP-9,and SMYD2.The relationship between LINC01106 or SMYD2 and miR-372-3p was confirmed by dual luciferase reporter assay.Results LINC01106 and SMYD2 were significantly up-regulated,and miR-372-3p was significantly down-regulated in liver cancer tissues and cell lines(P<0.05).Compared with those of blank control or transfected with si-NC,SMMC-7721 cells transfected with si-LINC01106 had significantly lower mRNA levels of LINC01106 and SMYD2,OD490 value,EdU-positive cell ratio,wound healing ratio,invasive cell number and protein levels of PCNA,MMP-9,MMP-2 and SMYD2,but higher expression level of miR-372-3p(P<0.05).Compared with those transfected with si-LINC01106+inhibitor-NC or si-LINC01106,SMMC-7721 cells transfected with si-LINC01106+miR-372-3p inhibitor had significantly higher mRNA levels of LINC01106 and SMYD2,OD490 value,EdU-positive cell ratio,wound healing ratio,invasive cell number and protein levels of PCNA,MMP-9,MMP-2 and SMYD2,but lower expression level of miR-372-3p(P<0.05).LINC01106 targeted a negative regulation of miR-372-3p,while miR-372-3p targeted a negative regulation of SMYD2.Conclusion Knockdown of LINC01106 inhibits migration,invasion,and proliferation of SMMC-7721 cells by targeting the miR-372-3p/SMYD2 axis.
作者
胡忠卓
刘红英
HU Zhongzhuo;LIU Hongying(Department of Gastroenterology,Pangang Group General Hospital,Sichuan,Panzhihua 617000,China)
出处
《河北医药》
2025年第5期721-727,共7页
Hebei Medical Journal
基金
宜宾市科技计划项目(编号:2021SF002)。
